玉米雄性不育突变体<i>x50</i>的基因定位与遗传分析

doi:10.7668/hbnxb.20193481

摘要/Abstract

摘要：

为了挖掘雄性不育种质资源,鉴定雄性育性基因,为玉米雄性不育化制种提供基础材料。以玉米雄性不育突变体x50为试验材料,研究突变体雄性不育表型,构建x50与自交系Mo17的F₁和F₂群体,确定突变体x50雄性不育性状的遗传模式。以F₂群体为材料,应用图位克隆技术定位雄性育性基因X50,通过基因等位性测验确定候选基因。结果显示,与野生型相比,雄性不育突变体x50花药不能从颖壳露出,花药体积较小且萎蔫,无成熟花粉粒形成。F₁群体植株均表现为雄性可育,F₂群体植株出现雄性育性分离,可育植株与不育植株分离比例符合3∶1,说明突变体x50不育性状受1对隐性核基因控制。通过图位克隆方法将雄性育性基因X50定位于玉米第2染色体分子标记2-4901与2-4963之间,物理区间为237.42~241.39 Mb。定位区间内候选基因分析发现,区间存在玉米雄性不育基因ZmMs33。以ms33纯合突变体ms33-6029和ms33-6052分别与x50杂合型+/x50杂交,杂交后代可育植株与不育植株分离比例符合1∶1,表明x50是ZmMs33基因一个等位突变体。玉米雄性不育突变体x50的鉴定为玉米杂交种子生产和ZmMs33基因功能研究提供了种质材料。

关键词: 玉米, 雄性不育基因X50, 基因定位, 花药, 等位性测验

Abstract:

In order to explore the germplasm resources of male sterile line and identify the male fertility genes, which would provide the basic materials for the maize seed production by the male sterile line. Using the male sterile mutant x50 as the experimental material,the male sterile phenotype of x50 was studied,and the F₁ and F₂ populations of x50 and inbred line Mo17 were constructed to determine the genetic pattern of the male sterile trait in x50.With the F₂ population,the male fertility gene X50 was identified by the map-based cloning.Furthermore,the candidate gene was confirmed by the allelism test.The results showed that compared with the wild type,the x50 anthers did not emerge from the glume,and was smaller and wilted,and no mature pollen grains were formed.All F₁ plants were fertile,and F₂ plants displayed 3∶1 segregation ratio between fertile and sterile plants,indicating that the monofactorial recessive inheritance of x50.The gene X50 was mapped to the interval from the molecular marker 2-4901 to 2-4963 on chromosome 2 with a physical range of 237.42 to 241.39 Mb by map-based cloning.Candidate gene analysis found that the male fertility gene ZmMs33 was located in the mapping region.In addition,the test crosses of the ms33 mutants(ms33-6029 and ms33-6052)and heterozygous +/x50 revealed 1∶1 segregation ratio between fertile and sterile plants.These results suggested that x50 was an allele mutant of ZmMs33 gene.The identification of the male sterile mutant x50 provided germplasm resources for maize hybrid seed production and functional study of ZmMs33 gene.

Key words: Maize, Male sterile gene X50, Gene mapping, Anther, Allelism test

郭瑶晴, 孙晓靖, 连玉杰, 陈慧, 孙华越, 张雪海, 汤继华, 陈晓阳. 玉米雄性不育突变体x50的基因定位与遗传分析[J]. 华北农学报, 2023, 38(1): 17-22. doi: 10.7668/hbnxb.20193481.

GUO Yaoqing, SUN Xiaojing, LIAN Yujie, CHEN Hui, SUN Huayue, ZHANG Xuehai, TANG Jihua, CHEN Xiaoyang. Gene Mapping and Genetic Analysis of Male Sterile Mutant x50 in Maize[J]. Acta Agriculturae Boreali-Sinica, 2023, 38(1): 17-22. doi: 10.7668/hbnxb.20193481.

http://www.hbnxb.net/CN/Y2023/V38/I1/17

图/表 7

图1 野生型和突变体x50表型比较 A.野生型(WT)和x50植株;B.野生型(WT)和x50雄穗;C.野生型(WT)和x50小穗;D,E.野生型(WT,D)和x50(E)成熟花粉粒。

Fig.1 Phenotypic comparison between the wild type and x50 mutant A.The plants of the wild type and x50;B.The tassels of the wild type and x50;C.The spikelets of the wild type and x50;D,E.The mature pollen grains of the wild type(D)and x50(E).

表1 F₂群体雄性育性表型分析

Tab.1 Phenotypic analysis of male fertility in F₂ population

时间和地点 Time and location	可育株数 No. of fertile plants	不育株数 No. of sterile plants	χ² (3∶1)
2019年,海口 2019,Haikou	153	48	0.13
2019年,原阳 2019,Yuanyang	190	64	0.01

图2 X50基因连锁标记的鉴定

Fig.2 The identification of linked markers with X50 gene

表2 X50基因定位引物

Tab.2 Primers used in the gene mapping of X50

引物名称 Primer name	引物序列(5'—3') Primer sequence(5'—3')
2-4791-F	GTCGCTGCTGCCCATGTTATTC
2-4791-R	TGCTGTGCCCGTGATCTCATC
2-4901-F	AACTCCAATCCGCCTCGACTG
2-4901-R	GTAGCCGTATCCTCGTCGTCTG
2-4963-F	TGCTGGTTAATCTGTGCCTTGC
2-4963-R	ACCTGTGTTGCCAAACTTGTTG

图3 X50基因定位

Fig.3 The gene mapping of X50

图4 突变体x50和ms33等位性测验 A.ms33-6029与杂合型+/x50杂交后代突变型(x50/ms33-6029)和野生型(WT)雄穗;B和C.ms33-6029与杂合型+/x50杂交后代突变型(x50/ms33-6029,B)和野生型(WT,C)成熟花粉粒。

Fig.4 Allelism test of x50 and ms33 mutant A.The tassels of x50/ms33-6029 and wild type derived from the cross of ms33-6029 and heterozygous+/x50;B and C.The mature pollen grains of x50/ms33-6029 and wild type derived from the cross of ms33-6029 and heterozygous+/x50.

表3 ms33突变体与杂合型+/x50杂交后代育性表型

Tab.3 Fertility phenotype of the crosses of ms33 mutant and heterozygous +/x50

组合 Combinations	不育株数 No. of sterile plants	可育株数 No. of fertile plants	χ² (1∶1)
ms33-6029×+/x50	34	28	0.58
ms33-6052×+/x50	16	17	0.03

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玉米雄性不育突变体x50的基因定位与遗传分析

Gene Mapping and Genetic Analysis of Male Sterile Mutant x50 in Maize

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